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A continuous cuticle is required for normal development of Arabidopsis embryos. Despite this barrier, embryos remain sensitive to outside signals. In this issue of Developmental Cell, De Giorgi et al. show that the embryonic cuticle is relatively permeable and becomes sealed during germination in response to peptide signaling.

In flowering plants, the asymmetrical division of the zygote is the first hallmark of apical-basal polarity of the embryo and is controlled by a MAP kinase pathway that includes the MAPKKK YODA (YDA). In Arabidopsis, YDA is activated by the membrane-associated pseudokinase SHORT SUSPENSOR (SSP) through an unusual parent-of-origin effect: SSP transcripts accumulate specifically in sperm cells but are translationally silent. Only after fertilization is SSP protein transiently produced in the zygote, presumably from paternally inherited transcripts. SSP is a recently diverged, Brassicaceae-specific member of the BRASSINOSTEROID SIGNALING KINASE (BSK) family. BSK proteins typically play broadly overlapping roles as receptor-associated signaling partners in various receptor kinase pathways involved in growth and innate immunity. This raises two questions: How did a protein with generic function involved in signal relay acquire the property of a signal-like patterning cue, and how is the early patterning process activated in plants outside the Brassicaceae family, where SSP orthologs are absent? Here, we show that Arabidopsis BSK1 and BSK2, two close paralogs of SSP that are conserved in flowering plants, are involved in several YDA-dependent signaling events, including embryogenesis. However, the contribution of SSP to YDA activation in the early embryo does not overlap with the contributions of BSK1 and BSK2. The loss of an intramolecular regulatory interaction enables SSP to constitutively activate the YDA signaling pathway, and thus initiates apical-basal patterning as soon as SSP protein is translated after fertilization and without the necessity of invoking canonical receptor activation.

Vascular plants have an open body plan and continuously generate new axes of growth, such as shoot or root branches. Apical-to-basal transport of the hormone auxin is a hallmark of every axis, and the resulting pattern of auxin distribution affects plant development across scales, from overall architecture to cellular differentiation. How the first axis is initiated in the early embryo is a long-standing question. While our knowledge is still sparse, some of the key players of axialization have emerged, and recent work points to specific models for connecting cellular polarity to the asymmetric division of the zygote and domain specific gene expression to the organization of basipetal auxin flux.

Summary In flowering plants, developing embryos reside in maternal sporophytes. It is known that maternal generation influences the development of next-generation embryos; however, little is known about the signaling components in the process. Previously, we demonstrated that Arabidopsis mitogen-activated protein kinase 6 (MPK6) and MPK3 play critical roles in plant reproduction. In addition, we noticed that a large fraction of seeds from mpk6 single-mutant plants showed a wrinkled seed coat or a burst-out embryo phenotype. Here, we report that these seed phenotypes can be traced back to defective embryogenesis. The defective embryos have shorter suspensors and reduced growth along the longitudinal axis. Furthermore, the cotyledons fail to bend over to progress to the bent-cotyledon stage. As a result of the uneven circumference along the axis, the seed coat wrinkles to develop raisin-like morphology after dehydration. In more severe cases, the embryo can be pushed out from the micropylar end, resulting in the burst-out embryo seed phenotype. Genetic analyses demonstrated that the defective embryogenesis of the mpk6 mutant is a maternal effect. Heterozygous or homozygous mpk6 embryos have defects only in mpk6 homozygous maternal plants, but not in wild-type or heterozygous maternal plants. The loss of function of MKK4/MKK5 also results in the same phenotypes, suggesting that MKK4/MKK5 might act upstream of MPK6 in this pathway. The maternal-mediated embryo defects are associated with changes in auxin activity maxima and PIN localization. In summary, this research demonstrates that the Arabidopsis MKK4/MKK5–MPK6 cascade is an important player in the maternal control of embryogenesis.

SummaryThe cell types of the plant root are first specified early during embryogenesis and are maintained throughout plant life. Auxin plays an essential role in embryonic root initiation, in part through the action of the ARF5/MP transcription factor and its auxin-labile inhibitor IAA12/BDL. MP and BDL function in embryonic cells but promote auxin transport to adjacent extraembryonic suspensor cells, including the quiescent center precursor (hypophysis). Here we show that a cell-autonomous auxin response within this cell is required for root meristem initiation. ARF9 and redundant ARFs, and their inhibitor IAA10, act in suspensor cells to mediate hypophysis specification and, surprisingly, also to prevent transformation to embryo identity. ARF misexpression, and analysis of the short suspensor mutant, demonstrates that lineage-specific expression of these ARFs is required for normal embryo development. These results imply the existence of a prepattern for a cell-type-specific auxin response that underlies the auxin-dependent specification of embryonic cell types.

Arabidopsis embryos follow a predictable sequence of cell divisions, facilitating a genetic analysis of their early development. Both asymmetric divisions and cell-to-cell communication are probably involved in generating specific gene expression domains along the main axis within the first few division cycles. The function of these domains is not always understood, but recent work suggests that they may serve as a basis for organizing polar auxin flux. Auxin acts as systemic signal throughout the life cycle and, in the embryo, has been demonstrated to direct formation of the main axis and root initiation at the globular stage. At about the same time, root versus shoot fates are imposed on the incipient meristems by the expression of antagonistic regulators at opposite poles of the embryo. Some of the key features of the embryonic patterning process have emerged over the past few years and may provide the elements of a coherent conceptual framework.

SummaryDivision of the Arabidopsis zygote defines two fundamentally different developmental domains, the proembryo and suspensor. The resulting boundary separates domain-specific gene expression, and a signal originating from the proembryo instructs the suspensor to generate the root stem cell niche. While root induction is known to require the phytohormone auxin and the Auxin Response Factor MONOPTEROS, it has remained largely elusive how the two domains involved in this process are initially specified. Here, we show that the GATA factor HANABA TARANU (HAN) is required to position the inductive proembryo boundary. Mutations in HAN cause a coordinated apical shift of gene expression patterns, revealing that HAN regulates transcription in the basal proembryo. Key auxin transporters are affected as early as the 8 cell stage, resulting in apical redistribution of auxin. Remarkably, han embryos eventually organize a root independent of MONOPTEROS and the suspensor around a new boundary marked by the auxin maximum.

The primary axis and main tissue types of Arabidopsis are laid down in the early embryo. Apical-to-basal auxin flux functions as a global organizer of the axis, and recent reports are clarifying our mechanistic understanding of how a graded auxin distribution is generated and interpreted. Polar targeting of PIN transporters in the cells of the embryo is dynamic and linked to their phosphorylation status, suggesting a flexible mechanism for regulating auxin flux in space and time. PLETHORA transcription factors then interpret the graded auxin distribution to provide positional values along the axis in a dose-dependent manner. A comparable framework for tissue patterning in the radial dimension is still lacking, although cell surface signaling probably plays a key role.

Early embryonic development in the flowering plant Arabidopsis thaliana follows a predictable sequence of cell divisions. Anatomical hallmarks and the expression of marker genes in dynamic patterns indicate that new cell fates are established with virtually every round of mitosis. Although some of the factors regulating these early patterning events have been identified, the overall process remains relatively poorly understood. Starting at the globular stage, when the embryo has approximately 100 cells, the organization of development appears to be taken over by programs that regulate postembryonic patterning throughout the life cycle.

The Arabidopsis zygote divides asymmetrically into an embryonic apical cell and a basal cell with mostly extra-embryonic fate. This fundamental asymmetry sets the stage for further embryonic development, but the events mediating it are poorly understood. We have identified a MAPKK kinase gene, named YODA, that promotes extra-embryonic cell fates in the basal lineage. In loss-of-function mutants, the zygote does not elongate properly, and the cells of the basal lineage are eventually incorporated into the embryo instead of differentiating the extra-embryonic suspensor. Gain-of-function alleles cause exaggerated growth of the suspensor and can suppress embryonic development to a degree where no recognizable proembryo is formed. Our results imply that a MAP kinase cascade acts as a molecular switch promoting extra-embryonic fate.

Apical-to-basal auxin flux is a defining feature of land plants and determines their main body axis. How is the axis first set up in the embryo? Recent studies reveal that the establishment of embryonic polarity with the asymmetric first division as well as the separation of shoot and root fates within the proembryo depend on transcriptional regulation in the zygote and early embryo. Although the functional connections need to be better defined, this transcriptional network likely provides the positional information required for initiating the machinery capable of processing the systemic signal auxin in a context-dependent manner.

Distinct forms of cytokinesis characterise specific phases of development in plants. In Arabidopsis, as in many other species, the endosperm that nurtures the embryo in the seed initially develops as a syncytium. This syncytial phase ends with simultaneous partitioning of the multinucleate cytoplasm into individual cells, a process referred to as cellularisation. Our in vivo observations show that, as in cytokinesis, cellularisation of theArabidopsis endosperm is coupled to nuclear division. A genetic analysis reveals that most Arabidopsis mutations affecting cytokinesis in the embryo also impair endosperm cellularisation. These results imply that cellularisation and cytokinesis share multiple components of the same basic machinery. We further report the identification of mutations in a novel gene, SPÄTZLE, that specifically interfere with cellularisation of the endosperm, but not with cytokinesis in the embryo. The analysis of this mutant might identify a specific checkpoint for the onset of cellularisation.

The embryo of the flowering plant Arabidopsis develops by a regular pattern of cell divisions and cell shape changes. Mutations in the KNOLLE (KN) gene affect the rate and plane of cell divisions as well as cell morphology, resulting in mutant seedlings with a disturbed radial organization of tissue layers. At the cellular level, mutant embryos are characterized by incomplete cross walls and enlarged cells with polyploid nuclei. The KN gene was isolated by positional cloning. The predicted KN protein has similarity to syntaxins, a protein family involved in vesicular trafficking. During embryogenesis, KN transcripts are detected in patches of single cells or small cell groups. Our results suggest a function for KN in cytokinesis.

During embryogenesis, a single cell gives rise to different cell types, tissues and organs which are arranged in a biologically meaningful context, or pattern. The resulting basic body organization of higher plants, which is expressed in the seedling, provides a reference system for postembryonic development during which the meristems of the shoot and the root produce the adult body. The seedling may be viewed as the superimposition of two patterns: one along the apical-basal axis of polarity and the other perpendicular to the axis. To analyse mechanisms underlying pattern formation in the embryo, a genetic approach has been taken inArabidopsis. Mutations in a small number of genes alter one or the other of the two patterns. The mutant phenotypes suggest that early partitioning of the axis is followed by region-specific development, including the formation of the primary shoot and root meristems. The cloning of two genes involved in pattern formation provides a basis for mechanistic studies of how cells adopt specific fates in the developing embryo.

The Arabidopsis embryo establishes polarity and main tissue types with the first five rounds of cell division. In this issue of Developmental Cell, Gooh et al. (2015) provide tools toward elucidating this poorly understood process through the first movies and targeted manipulations of early embryos developing inside cultured seeds.